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. 2015 Dec 11;5:17935. doi: 10.1038/srep17935

Figure 3. Genipin inhibits autophagy-dependent inflammasome activation.

Figure 3

(A) Analysis of the cell death phenotype by Annexin-V/PI staining. LPS-primed BMDMs were pretreated with genipin and then stimulated with nigericin or Salmonella. The cells were stained with Annexin-V and PI. The percentage of cells positive for both Annexin V and PI is shown. (B) LPS-primed BMDMs were incubated with genipin and/or 5 μM wortmannin for 1 h and then stimulated with nigericin or Salmonella. The cells were fixed, permeabilized and stained for LC3. LC3 is shown in green, and cell nuclei are shown in blue (DAPI). (C,D) LPS-primed BMDMs were incubated with genipin and/or wortmannin and then stimulated with nigericin or Salmonella. The cell lysates were immunoblotted for LC3, β-tubulin or GAPDH. (E) LPS-primed BMDMs were incubated with genipin and/or wortmannin and then stimulated with nigericin. The cross-linked pellets were immunoblotted for ASC. (F,G) LPS-primed BMDMs were incubated with genipin and/or wortmannin and then stimulated with nigericin or flagellin. IL-1β secretion was measured by ELISA. (H) BMDMs were transfected with either siRNA targeting ATG5 or scrambled siRNA (NEG). Forty-eight hours after transfection, the cells were primed with LPS for 4 h and then stimulated with genipin before nigericin or Salmonella challenge. IL-1β secretion was measured by ELISA. (I) LPS-primed BMDMs were incubated with the indicated doses of torkinib or rapamycin for 1 h and then stimulated with nigericin. IL-1β secretion was measured by ELISA. The data are from three independent experiments conducted in triplicate. *P < 0.05 and **P < 0.01 compared with controls.